Klenow Fragment

  • Thread starter rockind78
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Main Question or Discussion Point

I know what it is, but my question is: What are some (specifically) popular applications? I am guessing some sort of specialized PCR??? Any thoughts would be appreciated.
 

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  • #2
iansmith
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I know we use it to double strand DNA for electrophoretic mobility shift assay (EMSA) and DNA footprintings.

From the following site
http://arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/enzymes/klenow.html [Broken]

1. Synthesis of double-stranded DNA from single-stranded templates
2. Filling in recessed 3' ends of DNA fragments
3. Digesting away protruding 3' overhangs
4. Preparation of radioactive DNA probes
 
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Thank you Ian, that is just what I wanted to know!! :smile: This brings me to my next question I guess, being, what advantage would there be in utilizing JUST klenow fragment in synthesis of DS DNA fragments from SS DNA versus DNA polymerase III?
 
  • #4
iansmith
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Klenow fragment can start synthesis from a primed single strand whereas DNA Pol III requires ssb protein in addition to a primed ss.
 

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